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[淋巴瘤免疫分型] 颗粒酶和穿孔素

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发表于 2014-5-5 21:28:28 | 显示全部楼层 |阅读模式
悬赏1流星未解决
大家谁做过颗粒酶和穿孔素 ,具体用在什么病和临床意义能讲解一下么?如果我用一般的破膜胞内染色的方法对么?

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发表于 2014-5-5 21:39:15 | 显示全部楼层
主要是涉及细胞免疫占主导的疾病,如病毒和胞内菌感染等。一般的破膜胞内染色就满足要求了!最好是用荧光强度高的染料标记的。
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发表于 2014-5-5 21:46:35 | 显示全部楼层
颗粒酶和穿孔素常表达于细胞毒T和NK细胞中。给你两段英文介绍:

Perforin
Perforin, also known as cytolysin or C9RP (complement 9 related protein), is a 70 kD protein structurally similar to complement C9 component, and it is encoded by a gene located on the long arm of chromosome 10 . Once secreted on the target cell, perforin perforates the membrane and forms pores or channels  that enable the granzyme molecules simultaneously secreted to enter the target.
In steady state, perforin can be detected in NK cells , gamma/delta TCR T cells , and the subset of alpha/beta TCR T cells expressing CD11b . After in vitro activation with anti-CD3 and IL-2, perforin expression increases and the molecule becomes detectable also in some 50% of CD8+ CD11b– T cells, and in a small subset of CD4+ lymphocytes . Perforin is undetectable in resting CD34+ bone marrow cells, but it is present in the CD34+ KG1a cell line. It is also expressed in peripheral CD34+ cells after mobilization with G-CSF and chemotherapy but not with G-CSF alone .

Granzymes
Granzymes are a family of serine proteases stored in the granules of cytotoxic T lymphocytes and NK cells , which are able to induce apoptosis in target cells through activation of the caspase pathway Granzymes A, B and M are of interest in the diagnosis of hematological malignancies. These proteases differ according to their molecular binding sites, namely (i) arginine and lysine , (ii) aspartic acid , and (iii) methionine, leucine and norleucine , respectively.
Granzymes A and B are proteins of about 28–29 kD and are encoded by genes situated on the long arm of chromosome 5  and chromosome 14 , respectively. In resting conditions, granzyme positive cells are detectable in red pulp of the spleen, in tonsils, lymph nodes, liver, thymus and among lamina propria lymphocytes of the gut . In steady state, peripheral blood granzyme is detectable in CD3– CD16+ CD56+ NK cells only . According to some authors, granzymes A and B are equally distributed , while according to others the granzyme type distribution may depend on cell type and functional state . Intraepithelial gut T cells may express granzyme A in steady state, but also co-express granzyme B upon activation . Plasmacytoid dendritic cells have been demonstrated to serve as a source of granzyme B physiologically able to switch off T cell proliferation .
Some authors have postulated the presence of granzymes A and B and perforin in neutrophils , but others deny this fi nding .
Like perforin, granzyme B is undetectable in resting CD34+ bone marrow cells, but is present in the CD34+ KG1a cell line. It is also expressed in peripheral CD34+ cells after mobilization with G-CSF and chemotherapy but not with G-CSF alone .
Granzyme M is usually detectable in NK cells, gamma/ delta TCR T Lymphocytes and the small T CD56+ cell subset .


至于染色方法,常规的胞内染色方法即可。
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 楼主| 发表于 2014-5-5 21:58:28 | 显示全部楼层
谢谢,   
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发表于 2014-5-8 00:57:09 来自手机 | 显示全部楼层
我建议你用CD107a,这是一个与颗粒酶分泌一致的膜抗原。很好用

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组织样本处理不好?流式中文网原研的魔滤®魔杵®套装,低成本解决,高质量收获
发表于 2014-5-9 01:02:39 | 显示全部楼层
昨天用不了电脑,今天把文献补上
在线免费阅读:https://www.flowcyto.cn/bbs/forum.php?mod=attachment&aid=NTcxOXxjZjIzOWJkMnwxNzExNzExMjg1fDB8

下载PDF原文(需1颗流星): CD107 AND GrB.pdf (599.31 KB, 下载次数: 109, 售价: 1 流星)

组织样本处理不好?流式中文网原研的魔滤®魔杵®套装,低成本解决,高质量收获
 楼主| 发表于 2014-5-11 22:32:06 | 显示全部楼层
yuemin5 发表于 2014-5-9 01:02
昨天用不了电脑,今天把文献补上
在线免费阅读:

谢谢,已经下载下来
组织样本处理不好?流式中文网原研的魔滤®魔杵®套装,低成本解决,高质量收获
发表于 2014-5-12 20:07:48 | 显示全部楼层
很好的帖子
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发表于 2018-7-18 09:03:37 来自手机 | 显示全部楼层
niwanmao 发表于 2014-5-5 21:46
颗粒酶和穿孔素常表达于细胞毒T和NK细胞中。给你两段英文介绍:

Perforin

如果本身细胞是刺激扩增的,那么是不是就不需要刺激了,那是否需要抑制转运呢?
流式中文网FlowGuard®流式专用保存液,无需冻存,稳定保护各类流式样本,从容完成实验
发表于 2018-7-18 14:40:15 | 显示全部楼层
刘桂欢 发表于 2018-7-18 09:03
如果本身细胞是刺激扩增的,那么是不是就不需要刺激了,那是否需要抑制转运呢? ...

目前临床上是直接测的。
通过刺激扩增NK细胞,我没做过,但从原理上推测,最好是加。仅供参考。
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